Technique / Cell Biology / Signal transduction methods / In vitro Kinase Activity Assay

Kinase Assay Using In Vitro Translated Xenopus Mos Kinase

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By Radhika on 20-Mar-2008 Hello everyone i am doing kinase assay using P32. My fusion proteins are gst purified and then i incubate with nuclear extract to see if a particular mutant has a phosphorylation site. so i run the page, coomassie, destain my gel and dry it, then expose it. but my blot is all wierd and black as if some free radioactivity is stuck on it. can someone tell me what to change? should i rinse my gel with water several times before immersing it into coomassie solution? will this affect my radioactively labelled proteins? Rating: n/a Reply